Solarbio Tartrate-Resistant Acid Phosphatase Stain Kit (TRAP Stain Kit)

Kit Components:

Uvod:

Acid phosphatase (ACP) is widely distributed in various tissues, primarily in lysosomes, serving as a marker enzyme. Tartrate-resistant acid phosphatase (TRAP), found mainly in human alveolar macrophages and leukemia spleen cells, is released into cell follicles rather than the blood. TRAP in blood predominantly originates from osteoclasts, allowing assessment of osteoclast function.

The Tartrate-Resistant Acid Phosphatase (TRAP) Stain Kit principle involves hydrolysis of acid phosphatase with AS-BI substrate, releasing phosphoric acid and naphthol at acidic pH. A coupling of naphthol with diazo salts forms colored products localized in the cytoplasm. Positive staining indicates tartaric acid-resistant activity. It is commonly used for fresh blood smears, cell smears, and frozen sections.

Self-Provided Materials:

Distilled Water, Incubator, Slide, Microscope

Protocol (for reference only):

For Blood or Cell Smear:

1. Dry smear, fix in TRAP Fixative at 4°C for 30s-3mins.
2. Wash with distilled water, and air dry.
3. Apply TRAP Incubation Solution, and incubate at 37°C in the dark for 45-60min. Then wash.
4. Re-dye with Hematoxylin Solution or Methyl Green Solution for 2-3min.
5. View sections under the microscope after washing or sealing.

For Frozen Section:

1. Restore the section to 37°C by immersion in water for 1-2min.
2. Air dry, fix in TRAP Fixative for 1-3mins at 4°C.
3. Wash with distilled water, and air dry.
4. Apply TRAP Incubation Solution, and incubate at 37°C in the dark for 45-60min. Then wash.
5. Re-dye with Hematoxylin Solution or Methyl Green Solution for 2-3min.
6. View sections under the microscope after washing or sealing.

For Paraffin Section:

1. Dewax and rehydrate sections in graded alcohol.
2. Wash with distilled water, and air dry.
3. Apply TRAP Incubation Solution, and incubate at 37°C in the dark for 45-60min. Then wash.
4. Re-dye with Hematoxylin Solution or Methyl Green Solution for 2-3min.

Proizlaziti:

Clinical Significance:

1. TRAP staining aids in diagnosing hairy cell leukemia.
2. Positive staining is observed in immature monocytes of acute leukemia.
3. T lymphocytes exhibit positive TRAP staining.
4. Gaucher cells show strong positive staining.

Bilješka:

1. TRAP Incubation Solution may lose effectiveness quickly.
2. Reduce exposure time for staining frozen sections.
3. Use fresh samples immediately for accurate results.
4. Store tissue in fixative at 4°C for no more than 24 sati.
5. Control paraffin temperature during embedding to preserve enzyme activity.
6. Ensure high-quality xylene to prevent decomposition.
7. Wear experimental clothing and disposable gloves for safety.